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Quantitative gas chromatography-mass spectrometry isomer-specific measurement of hydroxy fatty acids in biological samples and food as a marker of lipid peroxidation.

We have developed a capillary gas chromatography-mass spectrometry method for the quantitative analysis of individual positional isomers of monohydroxy fatty acids derived from linoleic, arachidonic, eicosapentaenoic, or docosahexaenoic acid. Peroxidation of a particular polyunsaturated fatty acid results already in a complex mixture of positional isomers of hydroperoxy and hydroxy fatty acids. Catalytic hydrogenation of lipid extracts produces stable saturated hydroxy lipids from the complex mixtures typical of oxidized biological samples, simultaneously simplifying the analytical problem and eliminating oxidation artifacts. After saponification and methylation, monohydroxy fatty acid methyl esters are purified by solid-phase extraction and partially resolved using a CP Sil 19 column following on-column derivatization of the hydroxy groups with tetramethylammonium hydroxide. The resulting methoxy fatty acid methyl esters are subjected to electron impact mass spectroscopy. Two characteristic ions are produced for each positional isomer. Quantitative measurements were achieved by using odd chain C17 and C19 monohydroxy fatty acids as internal standards. The limit of detection of individual hydroxy fatty acid isomers is dependent on the total number of ions monitored. Monitoring 11 pairs of ions simultaneously gives limits of detection of 10 ng. Sensitivity is much higher by monitoring fewer ions and as little as 0.2 ng of a single isomer can be detected. The method has been applied for the quantitative analysis of hydroxy (plus hydroperoxy) fatty acids in plasma, adipose tissue, oils, and foods. To date over 1000 samples have been analyzed using the method described in this paper.

Dietary Fats, Unsaturated↗

A monoclonal antibody-based ELISA for the analysis of the insecticide flucythrinate in environmental and crop samples.

A competitive enzyme-linked immunosorbent assay (ELISA) has been developed for the detection of the insecticide flucythrinate in environmental and food samples. Two types of haptens, the acid moiety that is the hydrolyzed product of flucythrinate, and the carboxylated propyl derivative of the alcohol moiety, were used to prepare monoclonal antibodies (MAbs). Five MAbs, which raised against the former hapten, were reactive with flucythrinate. Among them, MAb F1A27-4 showed the highest activity toward flucythrinate, and did not cross-react with other pyrethroids such as cycloprothrin, fenvalerate, fluvalinate, etofenprox and silafluofen. The assay conditions of indirect competitive ELISA with MAb F1A27-4 were studied to optimize the detection of flucythrinate in environmental and food samples. Incubation at 4 degrees C in the assay buffer, pH 8, with 300 mM sodium chloride improved the sensitivity. The addition of rabbit serum albumin or rabbit antiserum and the presence of 50 ml litre-1 of methanol reduced matrix effects of the samples. Under optimized conditions, the ELISA detected flucythrinate spiked in water, soil, and extracts of apple and tea samples down to 10 mg litre-1, 0.2 mg litre-1, 0.3 mg litre-1 and 0.3 mg litre-1, respectively. The mean recovery and CV ranged from 91% to 120% and from 5% to 12%, respectively. The ELISA results in apple samples correlated well with those from LC-MS analysis (r2 = 0.99, n = 12).

Animals↗

Determination of acrylamide in processed foods by LC/MS using column switching.

An LC/MS method was developed for the determination of acrylamide (AA) in processed or cooked foods. AA was extracted with a mixture of water and acetone from homogenized food samples after the addition of 13C-labeled acrylamide (AA-1-(13)C) as an internal standard. The extract was concentrated, washed with dichloromethane for defatting, and cleaned up on Bond Elut C18, PSA and ACCUCAT cartridge-columns, and then AA was determined by LC/MS in the selected ion recording (SIR) mode. For the LC/MS analysis, four LC columns were connected in-line and the flow of the mobile phase was switched according to a time-program. Monitoring ions for AA were m/z 72 and 55, and those for AA-1-(13)C were m/z 73 and 56. AA and AA-1-(13)C were determined without interference from the matrices in all samples. The recoveries of AA from potato chips, corn snack, pretzel and roasted tea spiked at the level of 500 ng/g of AA were 99.5-101.0% with standard deviations (SD) in the range from 0.3 to 1.6%. The limits of detection and quantification of the developed method were 9 and 30 ng/g for AA in samples, respectively. The method was applied to the analysis of AA in various processed or cooked food samples purchased from retail markets. High levels of AA were found in potato chips and French-fried potato (467-3,544 ng/g). Fried and sugar-coated dough cakes (karinto) contained 374 and 1,895 ng/g. Corn snacks contained 117-535 ng/g of AA. Roasted foods (such as roasted sesame seed, roasted barley (mugi-cha), roasted tea (hoji-cha), coffee beans and curry powder) contained 116-567 ng/g of AA. Foods made from fish, egg and meat contained lower levels of AA than the plant-based foods. Foods containing much water showed a tendency to have low levels of AA compared with dry foods. The proposed method was applicable to the analysis of AA in variety of processed foods.

Acrylamides↗

Rapid detection of microbial contamination in frozen vegetables by automated impedance measurements.

Automated impedance measurements can be used to rapidly assess whether a sample of frozen vegetables contains greater or less than 10(5) organisms per g. Microorganisms growing pureed food samples cause a change in the impedance of the medium when the organisms reach a threshold concentration of between 10(6) and 10(7) organisms per ml. Estimates of the concentration of microorganisms initially present in the food sample can be made by recording the time required for the organisms in the sample to replicate to threshold levels. In this study, the detection times for 357 samples of frozen vegetables were compared with standard plate counts for each sample. The agreement between the two methods in distinguishing samples containing more than 10(5) organisms per g was 92.6% for 257 assorted frozen vegetables and somewhat higher (93 to 96%) when separate cutoff times were used for each type of vegetable. The time required for analysis was about 5 h, compared to the 48 to 72 h required for standard plate counts.

Bacteria↗

Effects of cross-sectional area on human bite studied with raw carrot and surimi gel.

The effects of the cross-sectional area of food samples on bite force with molar teeth were investigated using raw carrots and surimi gels. We evaluated human bite force for food samples with different sizes between the upper and lower molars using a multiple-point sheet sensor and electromyography (EMG). The bite force curve and EMG clearly showed textural characteristics of the carrot and gel. In particular, the first peak in the bite curves corresponded to breaking point in the compression test. With increasing cross-sectional area of both foodstuffs, the bite force and contact area increased and the average stress to which the specimen was subjected (mean stress) tended to decrease, while the stress produced between the teeth and the specimen (active stress) did not change. Chewing rhythm and EMG activities were not greatly influenced by sample size. These findings suggest that higher bite force might cause difficulty in biting food with a larger cross-sectional area.

Bite Force↗

Estimation of sucrose esters (E473) in foods using gas chromatography-mass spectrometry.

A method was developed for estimating the content in foods of the emulsifying additive E473, sucrose esters of fatty acids. The analytical approach taken to estimate the complex mixtures that comprise this additive involved, selective solvent extraction of the intact esters using a mixture of tetrahydrofuran and ethyl acetate, alkaline hydrolysis of the esters to liberate sucrose, and then GC-MS measurement of the liberated sucrose using GC-MS after acidic hydrolysis to glucose and fructose and then silylation. The method was developed to aid future estimates of intake of this food additive. The method determines the total sucrose esters content of a food sample and does not attempt to discrimination between individual sucrose esters when present as a mixture in a food sample. A single (average) factor is used to convert the liberated sucrose content into sucrose ester content. The method was applied to analysis of eight different food types (including bakery wares, sugar confectionery, dairy product, margarine, meat pies and a sauce) spiked with 0.5-1% of a mixture of three sucrose esters that spanned the hydrophilic/lipophilic balance (HLB) range 1-16. The limit of quantification was around 50 mg kg-1, which is more than adequate for these additives. The analytical recovery rate was 73-106% with an average of 91%. The precision of the method (RSD) was 6-18% (n = 3-20 for each food type) with an average RSD of 11%. The main analytical uncertainty is the conversion factor used to express sucrose ester content from the amount of sucrose liberated. The method is also applicable to sucroglycerides (E474).

Animals↗

[Development and application of a method for isolating Escherichia coli O157:H7 in the city of Gualeguaychú].

Culture media, reagents, and commercial kits were compared on artificially contaminated food samples. The objective was to find an isolation method for Escherichia coli O157:H7 sensitive, specific and accessible in terms of cost, requirements of equipments and qualification of the analyst. The adopted scheme consisted in a selective enrichment at 42 degrees C during 18 to 24 h, using an appropriate medium, in accordance with the nature of the sample, followed by a step of immunomagnetic separation and simultaneous isolation on a chromogenic agar and MacConkey sorbitol agar with potassium tellurite and cefixime, during 18-24 h at 37 degrees C. The presumptive colonies were confirmed as E. coli O157 by serological and biochemical tests. Secondly, this methodology was applied to food samples, water, bovine gastric content and manure. A total of 410 samples were studied: 279 from meat, 54 milk and dairy products, 6 from vegetables, 27 water samples and 44 bovine gastric content and manure. The frequency of isolation of E. coli O157:H7 was of 3.9%. The phenotypic and genotypic characterization of the isolates was performed. A simple isolation methodology for E. coli O157 was developed, which proved accessible to food laboratories of lower complexity. This methodology allowed the detection of this pathogen in food and environmental samples in Gualeguaychú City. The role of water as vehicle of infection was also established. The strains harbored the same virulence factors as those recovered from human disease.

Animals↗

The detection of enterotoxins and toxic shock syndrome toxin genes in Staphylococcus aureus by polymerase chain reaction.

A simple polymerase chain reaction (PCR)-based procedure was developed for the detection of fragments of staphylococcal enterotoxins (SEs) SEA, SEB, SEC, SED, SEE, SEG, SEH, and SEI together with the toxic shock syndrome toxin (TSST-1) genes of Staphylococcus aureus. One hundred and twenty-nine cultures of S. aureus were selected, 39 of which were recovered from 38 suspected staphylococcal food-poisoning incidents. The method was reproducible, and 32 different toxin genotypes were recognized. The presence of SE genes was associated with S. aureus strains reacting with phages in group III, and the TSST-1 gene with phages in group I. There was a 96% agreement between the PCR results for detection of SEA-D and TSST-1 as compared with a commercial reverse passive latex agglutination assay for the detection of SEs from cultures grown in vitro. Enterotoxin gene fragments were detected in S. aureus cultures recovered from 32 of the 38 suspected staphylococcal food poisoning incidents, and of these, 17 were associated with SEE, SEG, SEH, and SEI in the absence of SEA-D. Simple PCR procedures were also developed for the detection of SE directly in spiked food samples, and this was most successfully achieved in mushroom soup and ham. Detection was less successful in three types of cheese and in cream. SEA or SEB were detected by enzyme-linked immunosorbent assay in three food samples (two of which were associated with food poisoning incidents) naturally heavily contaminated with S. aureus: the appropriate SEA or SEB gene fragments were detected directly in these three foods by PCR.

Animals↗

Inductively Coupled Plasma Spectrometry Determination of Marker Elements for Childhood Soil Ingestion

The methodology developed with inductively coupled plasma spectrometry for the determination of marker elements (Al, Si, Ti, Zr, Y, Cr, La, Ce, Cr, and Nd) for childhood soil ingestion in biological (feces), environmental (dust and soil), and food samples are described. The quality assurance-quality control protocol for sample preparation, especially for food, diaper, and fecal samples and for determination of marker elements are presented.

Journal Article↗

Microbiological quality of food in relation to hazard analysis systems and food hygiene training in UK catering and retail premises.

A meta-analysis of eight UK food studies was carried out to determine the microbiological quality of food and its relationship with the presence in food businesses of hazard analysis systems and food hygiene training. Of the 19,022 premises visited to collect food samples in these studies between 1997 and 2002, two thirds (66%) were catering premises and one third (34%) were retail premises. Comparison with PHLS Microbiological Guidelines revealed that significantly more ready-to-eat food samples from catering premises (20%; 2,511/12,703) were of unsatisfactory or unacceptable microbiological quality compared to samples from retail premises (12%; 1,039/8,462) (p < 0.00001). Three quarters (76%) of retail premises had hazard analysis systems in place compared with 59% of catering premises (p < 0.00001). In 87% of retail premises the manager had received some form of food hygiene training compared with 80% of catering premises (p < 0.00001). From premises where the manager had received no food hygiene training a greater proportion of samples were of unsatisfactory and unacceptable microbiological quality (20% retail, 27% catering) compared with premises where the manager had received food hygiene training (11% retail, 19% catering) (p < 0.00001). Where the manager of the premises had received food hygiene training, documented hazard analysis systems were more likely to be in place (p < 0.00001). Higher proportions of samples of unsatisfactory and unacceptable microbiological quality (17% retail, 22% catering) were from premises where there was no hazard analysis system in place compared to premises that had a documented hazard analysis system in place (10% retail, 18% catering) (p < 0.00001). Our meta-analysis suggests that the lower microbiological quality of ready-to-eat foods from catering premises compared with those collected from retail premises may reflect differences in management food hygiene training and the presence of a hazard analysis system. The importance of adequate training for food handlers and their managers as a pre-requisite for effective hazard analysis and critical control point (HACCP) based controls is therefore emphasised.

Allied Health Personnel↗

Determination of lead and cadmium in foods by anodic stripping voltammetry: I. Development of method.

Food samples are dry ashed at 500 +/- 50 degrees C with a 10% aqueous K2SO4 solution used as an ashing aid. The ashed sample is dissolved in 50 mL 2% HNO3. Anodic stripping voltammetry is used to determine lead and cadmium in a mixture of the sample solution and an acetate electrolyte at pH 4.3 +/- 0.3. The estimated quantitation limits, based on a 10 g food sample, are 0.005 ppm for cadmium and 0.010 ppm for lead.

Cadmium↗

Detection, identification and characterization of bacteriocin-producing lactic acid bacteria from retail food products.

Forty bacteriocin-producing (Bac+) lactic acid bacteria (LAB) were isolated from food samples purchased from retail supermarkets and local farms. Of the 40 Bac+ isolates, 18 were isolated from 85 food samples by enrichment (21% isolation rate) whereas eight were obtained from 63 samples by direct plating (13% isolation rate). By direct plating, Bac+ LAB were detected at levels up to 2.4 x 10(5) cfu/g in ready-to-eat meats. The Bac+ isolates were identified by carbohydrate fermentation patterns, SDS-PAGE protein patterns, and other biochemical characteristics; SDS-PAGE proved invaluable in identifying strains that could not be identified by other means. Differential inhibitory spectra against indicator microorganisms assisted in the identification of 19 unique Bac+ isolates. Bac+ LAB included Enterococcus faecalis, Lactobacillus curvatus, Lb. delbrueckii, Lb. plantarum, Lactococcus lactis, and Pediococcus acidilactici. Lb. curvatus (four strains) and Lc. lactis (nine strains) were the only isolates inhibitory to foodborne pathogens including Listeria monocytogenes, Bacillus cereus, Clostridium perfringens and Staphylococcus aureus. Some Lc. lactis isolates inhibited as many as nine Gram-positive genera. Lb. curvatus FS47 and FS65 grew to high cell densities and produced bacteriocin at 6 degrees C; however, Lc. lactis FS56 produced greater levels of bacteriocin at lower cell densities. The high incidence of Bac+ LAB detected in retail foods indicates that the public is consuming a wide variety of Bac+ LAB that occur as natural contaminants. These data suggest a greater role for bacteriocins as biopreservatives in food.

Animals↗

Automatic microgravimetric determination of fats in milk products by use of supercritical fluid extraction with on-line piezoelectric detection.

An on-line supercritical fluid extraction-piezoelectric detection system was developed and applied to the quantitative gravimetric determination of total fat in food samples (skimmed milk and cocoa). The proposed assembly provides all the advantages of an on-line system as regards automation, in addition to acceptable sensitivity and precision. Its strength lies in the design of the interface between the supercritical fluid extractor and the piezoelectric detector. Samples of skimmed milk and cocoa are weighed in the extraction thimble, previously loaded with I g of diatomaceous earth. A temperature of 100 degrees C and a CO2 fluid density of 0.60 mg/ml are used for extraction. The linear calibration range thus achieved is 0.005-0.07% w/w total fat, and the relative standard deviation is +/-2.3% (n=11; P=0.05). The throughput is six samples h(-1) (for the overall process). The proposed method was used to determine the total fat in food samples (milk, cocoa), the results being competitive with those of the Soxhlet methods for the same purpose.

Animals↗

[The preparation of food and environmental samples for residue analysis of pesticides and PCBs by continuous steam distillation].

The treatment of water, soil and food samples was tested with a modified equipment of glass for simultaneous steam distillation/solvent extraction. Spiked samples with 22 pesticides (most of them contained halogen) and PCB yielded good or very good recoveries after 1.5 h for water samples and after 5 h for soil and plant food samples, respectively. This method is ecologically beneficial, and because it needs only a small amount of solvent (less than 10 ml) it is not expensive and may be used as a quick screening in several fields.

Environmental Pollutants↗

Feasibility of immunodiagnostic devices for the detection of ricin, amanitin, and T-2 toxin in food.

Qualitative and quantitative comparisons were conducted of commercially available immunodiagnostic devices for the detection of three select agents with oral LD50 values > or = 0.1 mg/kg of body weight. Ricin (oral LD50 > 1 mg/kg), amanitin (oral LD50 approximately 0.1 mg/kg), and T-2 toxin (oral LD50 > 1 mg/kg) were spiked into beverages, produce, dairy, and baked goods and assayed using commercially available enzyme-linked immunosorbent assays (ELISAs) and lateral flow devices. In all cases, the commercial diagnostic kits successfully detected all three select agents at concentrations below what might be a health concern. The considerable difference between the limit of detection of the immunodiagnostic devices employed (typically < or = 0.020 microg/g) and the amount of the select agent necessary to pose a health threat in a single serving of food facilitated the design of protocols for the high throughput screening of food samples. These protocols entailed simple extraction methods followed by sample dilution. Lateral flow devices and sandwich ELISAs for the detection of ricin had no significant background problems due to the food matrices. Competitive ELISAs, which typically have unacceptably high background reactions with food samples, successfully detected amanitin and T-2 toxin.

Amanitins↗

Determination of cannabinoids in hemp food products by use of headspace solid-phase microextraction and gas chromatography-mass spectrometry.

A fully automated procedure using alkaline hydrolysis and headspace solid-phase microextraction (HS-SPME), followed by on-fiber derivatization and gas chromatographic-mass spectrometric (GC-MS) detection has been developed for determination of cannabinoids in hemp food samples. After addition of a deuterated internal standard, the sample was hydrolyzed with sodium hydroxide and submitted to direct HS-SPME. After absorption of analytes for on-fiber derivatization, the fiber was placed directly into the headspace of a second vial containing N-methyl- N-trimethylsilyltrifluoroacetamide (MSTFA), before GC-MS analysis. Linearity was good for Delta(9)-tetrahydrocannabinol (THC), cannabidiol, and cannabinol; regression coefficients were greater than 0.99. Depending on the characteristics of the matrix the detection limits obtained ranged between 0.01 and 0.17 mg kg(-1) and the precision between 0.4 and 11.8%. In comparison with conventional liquid-liquid extraction this automated HS-SPME-GC-MS procedure is substantially faster. It is easy to perform, solvent-free, and sample quantities are minimal, yet it maintains the same sensitivity and reproducibility. The applicability was demonstrated by analysis of 30 hemp food samples. Cannabinoids were detected in all of the samples and it was possible to differentiate between drug-type and fiber-type Cannabis sativa L. In comparison with other studies relatively low THC concentrations between 0.01 and 15.53 mg kg(-1) were determined.

Cannabinoids↗

A bacteriological study of some frozen and nonfrozen foods.

Over a period of 19 months, a total of 331 food samples were submitted to the Food Section of the Bacteriology Division for bacteriological examination. These included 184 samples of frozen seafoods from exporters and 147 samples of fresh, nonfrozen foods from food caterers. The total bacterial count for frozen seafoods ranged from 1 x 10(2) to 2.98 x 10(6) per gm with a mean of 2.14 x 10(5) per gm. Coliforms, Escherichia coli and Staphylococcus aureus were present in 48.9%, 3.3% and 8.2% of the samples examined respectively. Two of the cooked prawn samples showed the presence of Vibrio parahaemolyticus. For the fresh, nonfrozen foods, the total bacterial count ranged from 1 x 10(2) to 3.87 x 10(6) per gm with a mean of 2.58 x 10(5) per gm. The examination also showed that 74.8% were coliform positive, 14.9% were E. coli positive, and 4.8% were S. aureus positive. V. parahaemolyticus was not isolated in any of the samples tested. Other pathogens, namely, Vibrio cholerae, Salmonella and Shigella were not isolated from any of the foods examined. The bacterial levels in these foods were determined and their sanitary and public health significance is discussed.

Animals↗

Evaluation of the selenium content of the traditional Italian diet.

Food samples and ready-made meals from the traditional Italian-type diet were analysed for selenium content. The average Se content varied in food samples from 7 micrograms/kg w/w (fresh fruit) to 226 micrograms/kg w/w (fish). The highest average contents were obtained in the animal products and in legumes. Among ready-made foods the animal derived dishes were the richest in Se, representing 78% of the estimated total daily dietary intake of Se. The average daily dietary intake of selenium for Italian people is estimated to be 50.9 +/- 29.8 micrograms Se/day when results obtained on complete meals are used, while it is 45.0 +/- 30.8 micrograms Se/day when results on foods and statistical data on consumption are used.

Animals↗